三氧化二砷诱导K562/ADM细胞凋亡的作用机制研究

Arsenic trioxide-induced apoptosis in K562/ADM cells and its mechanisms

目的 :观察三氧化二砷 (As2 O3 )对K5 6 2 ADM细胞的诱导凋亡效应 ,探讨其作用机制。方法 :应用噻唑蓝 (MTT)比色法、Wright Giemsa染色、DNA琼脂糖凝胶电泳和流式细胞术 (FCM)观察K5 6 2 ADM细胞凋亡 ;FCM测定K5 6 2 ADM细胞Fas、Bcl 2、P5 3蛋白水平的变化 ;比色法检测Caspase3活性变化。结果 :As2 O3 可抑制K5 6 2 ADM细胞增殖 ;K5 6 2 ADM呈典型凋亡形态改变 ;DNA电泳可见梯状条带出现 ;FCM分析示亚G1期细胞比例增高 ,G2 M期阻滞 ;Fas、P5 3蛋白表达明显上调 ;Caspase3活性明显增强。结论 :As2 O3 可通过Fas依赖性Caspase3激活而诱导K5 6 2 ADM细胞凋亡。

AIM: To observe the apoptosis of K562/ADM cells induced by As_2O_3 and to explore its possible mechanisms. METHODS: MTT assay,Wright-Giemsa staining,DNA agarose gel electrophoresis and cell cycle analysis were used to examine apoptosis in K562/ADM cells. Expression levels of Fas, Bcl-2 and P53 antigens were measured with FCM. Colorimetric assay was employed to detect the activation of Caspase3. RESULTS: As_2O_3 inhibited growth of K562/ADM cells. Morphological changes typical of apoptosis were observed through light microscopy. Agarose gel electrophoresis showed evident DNA fragmentation. Cell cycle analysis indicated increased Sub-G_1 proportion and apoptosis rate,as well as apparent G_2/M phase arrest. The expression of Fas and P53 antigens significantly increased after application of As_2O_3. Caspase 3 was also activated by As_2O_3. CONCLUSION: As_2O_3 induces apoptosis in K562/ADM cells by activating Caspase 3 via a Fas-dependent pathway.